PMID- 29798616 OWN - NLM STAT- MEDLINE DCOM- 20180627 LR - 20211015 IS - 1002-1892 (Print) IS - 1002-1892 (Linking) VI - 31 IP - 4 DP - 2017 Apr 15 TI - [Effects of different mechanical stretch conditions on differentiation of rat tendon stem cells]. PG - 481-488 LID - 10.7507/1002-1892.201611102 [doi] AB - OBJECTIVE: To investigate the effects of different mechanical stretch conditions on the differentiation of rat tendon stem cells (TSCs), to find the best uniaxial cyclic stretching for TSCs tenogenic differentiation, osteogenic differentiation, and adipogenic differentiation. METHODS: TSCs were isolated from the Achilles tendons of 8-week-old male Sprague Dawley rats by enzymatic digestion method and cultured. The TSCs at passage 3 were randomly divided into 5 groups: group A (stretch strength of 4% and frequency of 1 Hz), group B (stretch strength of 4% and frequency of 2 Hz), group C (stretch strength of 8% and frequency of 1 Hz), group D (stretch strength of 8% and frequency of 2 Hz), and group E (static culture). At 12, 24, and 48 hours after mechanical stretch, the mRNA expressions of the tenogenic differentiation related genes [Scleraxis (SCX) and Tenascin C (TNC)], the osteogenic differentiation related genes [runt related transcription factor 2 (RUNX2) and distal-less homeobox 5 (DLX5)], and the adipogenic differentiation related genes [CCAAT-enhancer-binding protein-alpha (CEBPalpha) and lipoprteinlipase (LPL)] were detected by real-time fluorescent quantitative PCR and the protein expressions of TNC, CEBPalpha, and RUNX2 were detected by Western blot. RESULTS: The mRNA expressions of SCX and TNC in group B were significantly higher than those in groups A, C, D, and E at 24 hours after mechanical stretch ( P<0.05). The mRNA expressions of CEBPalpha and LPL in group D were significantly higher than those in groups A, B, C, and E at 48 hours after mechanical stretch ( P<0.05). The mRNA expressions of RUNX2 and DLX5 in group C were significantly higher than those in groups A, B, D, and E at 24 hours after mechanical stretch ( P<0.05). Western blot detection showed that higher protein expression of TNC in group B than group E at each time point after mechanical stretch ( P<0.05), and the protein expression of CEBPalpha was significantly inhibited when compared with group E at 24 hours after mechanical stretch ( P<0.05). At 24 hours after mechanical stretch, the protein expression of RUNX2 in group C was significantly higher than that in group E ( P<0.05); and the protein expression of TNC was significantly lower than that in group E at 24 and 48 hours after mechanical stretch ( P<0.05). At 48 hours after mechanical stretch, the protein expression of CEBPalpha was significantly increased and the protein expression of TNC was significantly decreased in group D when compared with group E ( P<0.05), but no significant difference was found in the protein expression of RUNX2 between groups D and E ( P>0.05). CONCLUSION: The mechanical strain could promote differentiation of TSCs, and different parameter of stretch will lead to different differentiation. The best stretch condition for tenogenic differentiation is 4% strength and 2 Hz frequency for 24 hours; the best stretch condition for osteogenic differentiation is 8% strength and 1 Hz frequency for 24 hours; and the best stretch condition for adipogenic differentiation is 8% strength and 2 Hz frequency for 48 hours. FAU - Li, Pao AU - Li P AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Gao, Shang AU - Gao S AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Zhou, Mei AU - Zhou M AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Tang, Hong AU - Tang H AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Mu, Miduo AU - Mu M AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Zhang, Jiqiang AU - Zhang J AD - Department of Neurobiology, Third Military Medical University, Chongqing, 400038, P.R.China. FAU - Tang, Kanglai AU - Tang K AD - Department of Orthopedics, Orthopedic Center of Chinese PLA, Southwest Hospital, Third Military Medical University, Chongqing, 400038, P.R.China.tangkanglai@hotmail.com. LA - chi PT - Journal Article PL - China TA - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi JT - Zhongguo xiu fu chong jian wai ke za zhi = Zhongguo xiufu chongjian waike zazhi = Chinese journal of reparative and reconstructive surgery JID - 9425194 SB - IM MH - Animals MH - *Cell Differentiation MH - Cells, Cultured MH - Male MH - Mesenchymal Stem Cells MH - *Osteogenesis MH - Rats MH - Rats, Sprague-Dawley MH - *Stem Cells MH - Tendons/*cytology PMC - PMC8498182 OTO - NOTNLM OT - Mechanical stretch OT - differentiation OT - rat OT - tendon stem cells EDAT- 2018/05/26 06:00 MHDA- 2018/06/28 06:00 PMCR- 2017/04/01 CRDT- 2018/05/26 06:00 PHST- 2018/05/26 06:00 [entrez] PHST- 2018/05/26 06:00 [pubmed] PHST- 2018/06/28 06:00 [medline] PHST- 2017/04/01 00:00 [pmc-release] AID - zgxfcjwkzz-31-4-481 [pii] AID - 10.7507/1002-1892.201611102 [doi] PST - ppublish SO - Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2017 Apr 15;31(4):481-488. doi: 10.7507/1002-1892.201611102.